KMID : 1377020160130040388
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Tissue Engineering and Regenerative Medicine 2016 Volume.13 No. 4 p.388 ~ p.395
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Inhibition of fibrotic contraction by C-phycocyanin through modulation of connective tissue growth factor and ¥á-smooth muscle actin expression
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An Eun-jin
Park Hyun-ju Ae-Ri Cho Lee
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Abstract
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The effects of C-phycocyanin (C-pc), a phycobiliprotein, on the expression of pro-fibrotic mediators in hyper-tropic scarring such as connective tissue growth factor (CTGF) and ¥á-smooth muscle actins (¥á-SMA) were investigated in relation to trans-differentiation of fibroblast to myo-fibroblast, an icon of scar formation. C-pc was isolated from Spirulina Platensis extract using sonication method and C-pc concentration was determined by Bennet and Bogorad equation. ¥á-SMA and CTGF levels in wounded primary human dermal fibroblasts were determined by western blot analysis and immuno-fluorescence confocal microscope was employed. Fibroblast contractility was examined by three-dimensional collagen lattice contraction assay. There was an elevation of ¥á-SMA (121%) and CTGF (143%) levels in wound cells as compared with non-wound cells. The does-response profiles of down regulation demonstrated that the maximum inhibitions of ¥á-SMA by 63% (p<0.05) and CTGF by 50% (p<0.1) were achieved by C-pc (6 nM) treated cells. In confocal assay, non-wound fibroblasts exhibited basal level of ¥á-SMA staining, while wounded cells without C-pc treatment showed strong up-regulation of ¥á-SMA by 147% (p<0.05). C-pc (6 nM) inhibited ¥á-SMA expression by 70% (p<0.05) and reduced collagen contraction by 29% (p<0.05). C-pc seemed to lessen the over expression of CTGF, ¥á-SMA, subsequently alleviating the fibrotic contracture. This study suggests the potential application of C-pc to regulation of the expression of pro-fibrotic mediators in scarring process and its potential usage as an efficient means for anti-fibrosis therapy.
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KEYWORD
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Phycocyanin, Wound fibrosis, Myofibroblast, ¥á-SMA, Connective tissue growth factor, Wound healing
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